• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.96-96
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• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• The Korean Journal of Malacology
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• v.27 no.3
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• pp.273-282
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• 2011

The ultrastructural characteristics of germ cells during spermatogenesis and mature sperm morphology in male Pinctada martensii were investigated by transmission electron microscope observation. The morphologies of the sperm nucleus and the acrosome of this species are the oval shape and cone shape, respectively. Spermatozoa are approximately $47-50{\mu}m$ in length including a sperm nucleus (about $1.24{\mu}m$ in length), an acrosome (about $0.60{\mu}m$ in length), and tail flagellum (about $45-47{\mu}m$). The axoneme of the sperm tail shows a 9+2 structure. In P. martensii in Pteriidae, a special substructure showing a thick and wide triangular shape which is composed of electron-dense opaque material (occupied about 50% of all, the upper part of the acrosomal vesicle), appeared in the upper region (part) of the acrosomal vesicle, while the lower region (part) of the acrosomal vesicle is composed of electron-lucent material. Thus, this special structure, which exist in the upper part of the acrosomal vesicle in P. martensii, is somewhat different from those of other subacrosomal vesicle in other families in subacrosomal vesicles. Therefore, we assume that the existence of a special substructure showing a thick and wide triangular shape in the acrosomal vesicle of the spermatozoon can be used as a key characteristic for identification of P. martensii or other species in Pteriidae in subclass Pteriomorphia. The number of mitochondria in the midpiece of the sperm of this species are five (exceptionally sometimes four), as one of common characteristics appear the same number of mitochondria in the same families of superfamilyies. This species in Pteriidae does not contain the axial rod and satellite fibres which appear in the species in Ostreidae in subclass Pteriomorphia. These characteristics can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or tools.